Methanolic extract of Ephedra alata inhibits breast cancer cells in vitro and in vivo

Objective:To determine the anticancer potential of the methanolic extract from Ephedra alata against breast cancer both in vitro and in vivo.Methods:The effects of the methanolic extract of Ephedra alata on the viability,migration as well as apoptosis of breast cancer 4T1 cells were measured using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay,Transwell assay,and annexin V-FITC staining assay,respectively.Histological examination was also carried out.Moreover,a murine breast cancer model was established to evaluate the inhibitory effect of the extract.Biochemical parameters including hepatic and non-hepatic enzymes,malondialdehyde,and glutathione were investigated.Results:The methanolic extract of Ephedra alata showed a strong anti-proliferative and anti-migratory activity against 4T1 cells in a dose-dependent manner.It also induced apoptosis in 4T1 cells.In an in vivo mouse model,the extract markedly inhibited tumor growth,reduced malondialdehyde,and hepatic and non-hepatic enzymes as well as increased glutathione level.Conclusions:The methanolic extract of Ephedra alata inhibits breast cancer in vitro and in vivo,which may be a promising anticancer agent.

TCM practitioners’attitudes and perceptions regarding the use of Ephedra sinica Stapf:An observational study

Objective:To understand the attitudes and perceptions of traditional Chinese medicine(TCM)practitioners in Beijing TCM hospitals regarding the use of Ephedra sinica Stapf(E.sinica,Ma Huang).Methods:A two-stage cross-sectional study was conducted using a questionnaire survey of TCM practi-tioners in Beijing TCM hospitals between April 2023 and March 2024.The questionnaire included de-mographic information,the clinical background of TCM practitioners,and the clinical application of E.sinica.Logistic regression analysis was used to analyze the relevant influencing factors when using E.sinica.Results:Of the 465 questionnaires collected,441 were valid.Among these,84.81%(374/441)reported having used E.sinica in clinical practice at least once.The commonly used doses of E.sinicadexcluding the pediatric departmentdwere 10 g for high doses,6 g for medium,and 3 g for low.The three most frequently used formulas for E.sinica included Maxing Shigan decoction,Mahuang decoction,and Xiao Qing Long decoction.The most common TCM patterns treated with a high dose of E.sinica were wind-cold exterior pattern,wind-cold invading the lung,and wind and water combat with meridians con-gealed by cold.The top three Western medical diagnoses when using E.sinica for treatment were common cold,pneumonia,and upper respiratory tract infections.Nearly half of the respondents reported experiencing adverse reactions from the oral administration of E.sinica,with the most common being palpitations,insomnia,and restlessness.Starting with a low dose and gradually increasing it as appro-priate was identified as an effective approach.Conclusion:This study investigated the attitudes and perceptions of TCM practitioners in Beijing TCM hospitals regarding the doseeefficacyeadverse reaction relationship of E.sinica,providing a reference for the safe and effective clinical use of E.sinica.

Preparation of Ephedra houttuynia Granule and Its Therapeutic Mechanism of Anti-Mycoplasma gallisepticum Infection Based on Network Pharmacology

The use of Chinese herbal medicines can replace antibiotics that cause drug-resistance problems,which are currently necessary for disease control.In this paper,a traditional Chinese medicine compound named Ephedra houttuynia granule for the treatment of Mycoplasma galliscepticum(MG)infection was prepared.Furthermore,its action mechanism was explored through network pharmacology.The optimal extraction and granulation processes of the compound were determined by high performance liquid chromatography(HPLC)method and L9 orthogonal test,and in the treatment experiment,Ephedra houttuynia granule showed a significant therapeutic effect on MG infection.In the study of network pharmacology,the results showed that the core targets of Ephedra houttuynia granule against MG infection were vascular endothelial growth factor(VEGFA),fos proto-oncogene(FOS),prepro-coagulation factor II(F2),etc.,the gene ontology/kyoto encyclopedia of genes and genomes(GO/KEGG)analysis results indicated that the signaling pathways of neuroactive ligand receptor interaction,cAMP,IL-17,T cell receptor,and tumor necrosis factor(TNF)might involve in anti-MG infection.In conclusion,this study would provide a new idea for elucidating the action mechanism of other diseases in veterinary clinic,which had a certain guiding significance.

蒙古族利用草麻黄（Ephedra sinica Stapf）资源的民族植物学研究

应用民族植物学的研究方法，记载和描述了蒙古族对草麻黄资源在经济利用、药物利用和文化利用方面的知识和经验，对民间利用的合理性和意义及草原地区草麻黄资源的开发和持续利用作了分析和探讨．

Ontogeny and Metamorphic Patterns of Female Reproductive Organs of Ephedra sinica Stapf (Ephedraceae)

Ontogeny and metamorphic. development of female reproductive organs in Ephedra sinica Stapf were surveyed. At the end of April, female cones began to initialize from the vegetative buds. Pollination occurred in mid-May and seeds matured at the beginning of July. The ontogenetic pattern of female reproductive organs of E. sin ca is basically similar to that of E. distachya L. The foliar nature of the outer envelope of the ovule in Ephedra is corroborated. Reduction of ovule number as a tendency of speciation in the genus is substantiated by the occurrence of tri-ovulate cones coupled with comprehensive characters of the genus. The metamorphic patterns as well as the leaf nature of the outer envelope indicate that female cones of Ephedra are compound while the female reproductive units of the ovulate cone have been reduced from secondary reproductive shoots similar to those of cordaites by phylogenetic transformation, fusion and reduction. Each fertile bract together with its axillary female reproductive unit composed the Bract Scale and Seed Scale Complex of Ephedra.

基于网络药理学与分子对接探讨麻黄附子细辛汤治疗老年支气管哮喘作用机制

目的:利用网络药理学与分子对接,探讨麻黄附子细辛汤治疗老年支气管哮喘的关键活性成分、核心靶点基因及作用机制。方法:通过TCMSP、SwissTargetsPrediction、UniProt数据库检索和筛选,获得麻黄附子细辛汤的活性成分及相关靶点;利用GeneCards、OMIM和Pharm GKB数据库搜索老年支气管哮喘的相关靶点,筛选出药物活性成分与疾病的交集靶点;通过STRING和Cytoscape软件创建PPI网络图;将疾病与中药活性成分的交集靶点导入至DAVID数据库进行GO和KEGG富集分析,以进一步探索潜在作用机制;使用AutoDock Vina软件进行分子对接,验证核心靶点与关键活性成分的结合能力以及稳定性。结果:获得麻黄附子细辛汤活性成分52种、活性成分靶点466个,疾病的相关靶点1399个,筛选药物与疾病的交集靶点217个;通过分析,确定麻黄附子细辛汤治疗老年支气管哮喘的关键活性成分为槲皮素、木犀草素、山柰酚等;核心靶点分别为肿瘤坏死因子、白细胞介素6、RAC-α丝氨酸/苏氨酸蛋白激酶等;KEGG富集分析主要涉及肿瘤坏死因子信号通路和PI3K/Akt等信号通路;分子对接结果显示核心靶点与关键活性成分结合活性良好。结论:基于网络药理学与分子对接得出麻黄附子细辛汤关键活性成分与肿瘤坏死因子、白细胞介素6等靶点相结合,并通过肿瘤坏死因子和PI3K/Akt等信号通路作用于老年支气管哮喘,为临床治疗及进一步研究提供理论依据。

A fossil-calibrated relaxed clock for Ephedra indicates an Oligocene age for the divergence of Asian and New World clades and Miocene dispersal into South America

Ephedra comprises approximately 50 species, which are roughly equally distributed between the Old and New World deserts, but not in the intervening regions （amphitropical range）. Great heterogeneity in the substitution rates of Gnetales （Ephedra, Gnetum, and Welwitschia） has made it difficult to infer the ages of the major divergence events in Ephedra, such as the timing of the Beringian disjunction in the genus and the entry into South America. Here, we use data from as many Gnetales species and genes as available from GenBank and from a recent study to investigate the timing of the major divergence events. Because of the tradeoff between the amount of missing data and taxon/gene sampling, we reduced the initial matrix of 265 accessions and 12 loci to 95 accessions and 10 loci, and further to 42 species （and 7736 aligned nucleotides） to achieve stationary distributions in the Bayesian molecular clock runs. Results from a relaxed clock with an uncorrelated rates model and fossil-based calibration reveal that New World species are monophyletic and diverged from their mostly Asian sister clade some 30 mya, fitting with many other Beringian disjunctions. The split between the single North American and the single South American clade occurred approximately 25 mya, well before the closure of the Panamanian Isthmus. Overall, the biogeographic history of Ephedra appears dominated by long-distance dispersal, but finer-scale studies are needed to test this hypothesis.

Identification of Two Cold Water-Soluble Polysaccharides from the Stems of Ephedra sinica Stapf

Two polysaccharides (ESP-A1 and ESP-A2) were isolated from the cold water extract of Ephedra sinica Stapf and purified through ethanol precipitation, deproteinization and by ion exchange and gel-filtration chromatography. Their molecular weight was determined using high performance size exclusion chromatog-raphy and evaporative light scattering detector (HPSEC-ELSD) and their monosaccharide composition was analyzed by high performance capillary electrophoresis (HPCE) based on pre-column derivatization with 1-phenyl-3-methyl-5-pyrazolone (PMP). It was shown that ESP-A1 consisted of xylose, arabinose, glucose, mannose and galactose and ESP-A2 consisted of xylose, arabinose, rhamnose and galactose, in a molar ratio (%) of 3.2: 61.1: 11.1: 12.9: 11.6 and 20.6: 67.7: 5.0: 6.7, respectively. The molecular weights (Mw) of ESP-A1 and ESP-A2 were 5.83 × 104 Da and more than 200 × 104 Da, respectively. To the best of our knowledge, two neutral polysaccharides are now being reported for the first time in this study.

Mechanism of improving skin barrier function in a mouse model of atopic dermatitis using Mahuang Lianqiao Chixiaodou decoction and its disassembled formula

Objective: To reveal the mechanism of Mahuang Lianqiao Chixiaodou decoction and its disassembled formula for improving the skin barrier function in a mouse model of atopic dermatitis(AD).Methods: Sixty specific-pathogen free male BALB/c mice were randomly divided into the control group,model group, whole formula group(WF), exterior-releasing formula group(ERF), interior-clearing formula group(ICF), and positive control group(PC). A mouse model of AD was established using the semiantigen 2,4-dinitrofluorobenzene induction method. The lesion scores, transepidermal water loss and p H, and skin histopathology of mice in each group were observed. The expressions of filaggrin, loricrin,and involucrin were detected by the streptavidin peroxidase immunohistochemical method and western blotting, and their mRNA expressions were detected by quantitative polymerase chain reaction.Results: Mice in the WF, ERF, ICF, and PC groups showed reduced skin lesion performance, improved histopathology, decreased skin lesion score, transepidermal water loss and pH, and upregulated expressions of proteins including filaggrin, loricrin, and involucrin, and their mRNAs. The most obvious regulatory effect was observed in the WF group, followed by the ICF, ERF, and PC groups, accordingly.Conclusions: Mahuang Lianqiao Chixiaodou decoction and its disassembled formula can improve the skin barrier function in a mouse model of AD by upregulating filaggrin, loricrin, and involucrin, and their mRNA expressions, and the most optimal effect was noted in the WF group, followed by the ICF and ERF groups, which suggests that the effect of clearing heat and resolving dampness in improving the skin barrier function of AD is more obvious and is one of the key treatments for AD.

Intervention of the Mahuang Lianqiao Chixiaodou decoction on immune imbalance in atopic dermatitis-like model mice

Objective:To explore the potential mechanism of intervention on the immune imbalance of atopic dermatitis(AD) by studying the effects of Mahuang Lianqiao Chixiaodou decoction(MLCD) on skin damage and inflammation factors in an AD-like mouse model.Methods:Ninety-six male BALB/c mice were divided into normal,model,positive control(mometasone furoate),and traditional Chinese medicine treatment(MLCD) groups by a random number table.2,4-dinitrofluorobenzene was used to induce AD-like mice in all groups except the normal group.The treatment or intervention was administered for seven consecutive days on days 4,18,32,and 39.The mRNA relative expressions of interleukin-4(IL-4),IL-10,interferon-γ(IFN-γ),thymic stromal lymphopoietin(TSLP),and the TSLP receptor(TSLPR) were measured using quantitative real-time polymerase chain reaction,and the serum immunoglobulin E,IL-4,IL-10,and IFN-γ levels were detected using enzyme-linked immunosorbent assay.Results:Compared with the normal group,the hematoxylin-eosin staining of the skin lesions of the mice in the model group was significantly thickened on days 11,25,and 39.Compared with the model group,the epidermal thickness of the positive control group was significantly alleviated on day 39(P <.001),and that of the MLCD group was significantly improved on days 25 and 39(P <.001).Compared with the four observation time points,MLCD had the best treatment effect on day 39 of the experiment and significantly improved the skin damage performance and relieved pathological lesions.On day 39,compared with the model group,MLCD downregulated the skin mRNA relative expressions of IL-4(P=.009),TSLP(P=.030),and TSLPR(P <.001),and reduced the mouse serum levels of IL-4(P=.003).For other serum indicators,no significant difference was observed between the model and MLCD groups.Conclusion:MLCD improved AD-like mice skin damage by regulating the Th1/Th2 immune imbalance.

Efficacy and safety of Mahuang Fuzi Xixin Decoction on sick sinus syndrome： Ameta – analysis

Objective： To evaluate the efficacy and safety of Mahuang Fuzi Xixin Decoction on sick sinus syndrome andprovide evidence for clinical practice. Methods： Randomized controlled trials of all the languages of MahuangFuzi Xixin Decoction on sick sinus syndrome were collected by computer search and manual retrieval. Theretrieval time was from January 2000 to January 2017. According to the inclusion and exclusion criteria, 2reviewers independently selected and extracted data, then evaluated the quality, cross-checked the information andevaluated the quality of menthodology. Through discussion or third reviewer to help solve the divergence, RevMan5.3 software was used to perform meta analysis. Results： A total of 7 documents （n = 612） were finally enrolled,with 358 in Mahuang Fuzi Xixin Decoction group （treatment group） and 254 in control group. Meta analysisshowed that the treatment （86.9%） was more effective than the control （70.1%）, the difference was statisticallysignificant （RR = 1.25, 95% CI：（1.15-1.37）, P 〈 0.001）; the treatment （17.0%） was safer than the control （49.8%）,the difference was statistically significant （RR=0.23,95% CI：（0.06-0.93）, P =0.04）. Conclusion： The existingclinical studies suggest that Mahuang Fuzi Xixin Decoction on sick sinus syndrome is effective and safe; due to thelimited quality of the enrolled documents, the above conclusions need more high-quality randomized controlledtrials to be verified.

Effects of ephedra asarum aconite decoction on Th1/Th2 balance in two animal models of allergic rhinitis

Objective:Ephedra asarum aconite decoction is used in the clinical treatment of allergic rhinitis.We investigated the effects of ephedra asarum aconite decoction on Th1/Th2 balance in two animal models of allergic rhinitis and in activated T cells in vitro.Methods:Animal models of simple and compound kidney yang deficiency types of allergic rhinitis were treated with ephedra asarum aconite decoction to observe the efficacy of this traditional Chinese medicine formula on allergic symptoms,serum cytokines,and tissue RNA levels.We also cultured and activated T cells from the mouse.After treatment with ephedra asarum aconite decoction,we tested the proliferation and differentiation of T cells.Results:Ephedra asarum aconite decoction relieved allergy symptoms in both types of animal models,and was more effective in the compound kidney yang deficiency model In addition to its anti-allergic efficacy,ephedra asarum aconite decoction significantly improved the health status of animals in the compound kidney yang deficiency groups,significantly made their bodyweight rising up,and significantly increased the T-bet mRNA/GATA-3 mRNA ratio.Moreover,ephedra asarum aconite decoction promoted the proliferation of activated T cells and inhibited the differentiation to Th2 cells in vitro.Conclusions:Ephedra asarum aconite decoction regulates the balance of Th1/Th2 to ameliorate allergic conditions.

Effects of the ephedra alkaloid methylephedrine on the basal evoked potential transportation and the long-term potentiation (LTP) in the rat hippocampal dentate granule cells in vivo

Objective The effect of the ephedra/ephedrine alkaloid methylephedrine(dl-methylephedrine hydrochloride for testing in this paper)on cognitive related synaptic plasticity was investigated by recording extracellular field evoked potentials and its LTP in hippocampal dentate granule cells in urethane-anaesthetized rats in vivo.Methods Single pathway recording of evoked field potentials was made from the dentate granule cells of hippocampal hemisphere in response to stimulation of the ipsilateral medial perforant path(MPP).Two parameters,the amplitude of population spike(PS amplitude)and the latency of the PS,were employed to evaluate the effects of drug on the overall changes in cellular responses.Results The present study show that methylephedrine 90 mg·kg-1 intraperitoneally,about 1/3 LD50,could increase the latency of the PS in hippocampal dentate granule cells by constant single stimulation of the MPP as the basal ransportation.However,the 30 mg·kg-1 and 10 mg·kg-1 dosage had no effect on the latency,and there are no influences of PS amplitude for all examinational groups.The methylephedrine 90 mg·kg-1 group significant enhanced the development of amplitude LTP in hippocampal dentate granule cells that induced by 60 Hz,60 pulses conditional tetanus in medial perforant path area.Also,the 30 mg·kg-1 group can promoted the maintenance of LTP induced by this tetanus,but no promotion on PS amplitude LTP appeared in this dosage and no any changes been found in 10 mg·kg-1 dosage group.Conclusions The ephedra/ephedrine alkaloid methylephedrine can modulate the synaptic plasticity in the lateral perforant path.A possible mechanism of methylephedrine on hippocampal LTP is been discussed.

Herbivory Effects on <i>Ephedra</i>spp. in the Chihuahuan Desert

Two species of Ephedra: E. trifurca and E. torreyana inhabit shrub and grassland habitats in the northern Chihuahuan Desert. E. torreyana is limited to black grama grasslands where grasses are taller than the shrub. E. torreyana is heavily browsed by vertebrates and E. trifurca is browsed during some years. We established an experiment with cylindrical exclosures that excluded rabbits and rodents, rabbits but accessible to rodents, for comparison with E. torreyana plants available to all herbivores. Plants accessible to all vertebrate herbivores were significantly smaller with shorter stem lengths than plants in exclosures. We concluded that E. torreyana in black grama grassland are largely hidden from vertebrate herbivores and that intense herbivory reflects the degraded state of the study site which makes the E. torreyana evergreen shrubs apparent to vertebrates.

Maintenance of Ephedra alata Seeds Viability via Storage Containers

Sequential incubation of seed samples yielded 20 fungal species belonging to 13 genera. The prevalent genera were Aspergillus (A. flavus and A. parasiticus), Fusarium (F. moniliforme and F. oxysporum) and Penicillium (Penicillium sp.). Such seedborne fungi differ in their colonization in different parts of seeds with most of them are colonized in seed coat, endosperm, and embryo of the seed. The usage of different storage containers for storing seeds indicated that the cotton cloth bags were the most favorable ones as they maintain seed moisture content (SMC) below the critical level resulting in minimum seed deterioration compared with other seed storage containers.

Action mechanism of Ephedra in treating allergic rhinitis based on network pharmacology

Objective:To study the mechanism of ephedra in treating allergic rhinitis by network pharmacology.Methods:the chemical components and corresponding targets of ephedra were searched and screened by TCMSP database.With"allergic rhinitis"as the key word,disease-related genes were searched in GeneCards and OMIM database.Using Venny2.1.0 mapping,Venn diagram and drug-disease common targets,network visualization software Cytoscape 3.6.1 track production elements-targets-disease network diagram,String database build their proteins interact with each other(PPI),and using R language to do gene ontology(GO)functional enrichment and pathway analysis and kyoto encyclopedia of genes and genome(KEGG)pathway enrichment analysis.Results:ephedrine was screened for 136 efficacy components and 88 common targets,and was predicted to intervene and regulate allergic rhinitis through AGE-RAGE signaling pathway in diabetic complications,Fluid shear stress and atherosclerosis,IL-17 signaling pathway,TNF signaling pathway and other signaling pathways in diabetic complications.Conclusion:preliminary prediction of the target and pathway of ephedra in the treatment of allergic rhinitis through network pharmacology can provide direction and evidence for the next experiment and clinic.

以麻黄为主药系列经方在儿童过敏性疾病辨治中的运用

以麻黄为主药系列经方在儿童过敏性疾病辨治中的运用。儿童过敏性疾病多年来是临床治疗的难点,以麻黄为主药系列方治疗该病临床疗效确切肯定。儿童过敏性疾病多因正气不足,肺、脾、肾功能低下,痰饮等伏邪宿根停留于体内,外邪引动而发病。麻黄为治疗咳喘之圣药,通过合理的配伍,麻黄与杏仁宣降相成,麻黄与石膏寒温并举,麻黄与附子温通表里。为了确保疗效,要重视遵循经方的原配伍比例、剂量、组方、煎服法和调护法。此外,还要明确麻黄类方及方证应用,即对方证,抓主证。

Mahuang Xixin Fuzi decoction protects against ovalbumin-induced allergic rhinitis by inhibiting type 2 innate lymphoid cells in mice

Objective:As a traditional herbal formula,Mahuang Xixin Fuzi decoction(MXFD)has been used to treat allergic rhinitis(AR).It regulates the transcription factor GATA3 in T cells,blocks Th2 skewing and rebalances the Th1/Th2 immunity.Type 2 innate lymphoid cells(ILC2s)are closely associated with GATA3.However,it remains unknown whether ILC2s could be one mechanism through which MXFD acts.We sought to elucidate the efficacy and mechanism of action of this decoction in AR treatment.Methods:Forty C57BL/6J female mice were equally divided into control,model,loratadine-and MXFDtreated groups in random.AR was induced by ovalbumin(OVA),and then treatment with loratadine or MXFD was administered.AR scores were evaluated and compared before and after treatment.Pathological changes in the nasal mucosa and lung were observed using hematoxylin-eosin staining of tissue samples.Activation of ILC2 in nasal mucosa was assessed by immunofluorescence and quantitative polymerase chain reaction analysis.ILC2 cell count in lungs was measured by flow cytometry and levels of interleukin-(IL)4,IL-5 and IL-13 in serum were detected by ELISA.Results:The decoction alleviated the symptoms of AR in mice,improved vascular congestion and expansion,glandular hyperplasia and inflammatory cell infiltration in mouse nasal mucosa and slowly improved the interstitial pneumonia and lesions.Meanwhile,MXFD reduced the number and percentage of ILC2s in the nasal mucosa and lungs,downregulated the expression of GATA3 mRNA and RORa mRNA,and reduced the related inflammatory cytokine levels,including IL-4,IL-5 and IL-13.Conclusion:These data suggest that inhibition of ILC2s by MXFD may be an important means by which to treat AR.

Study on the mechanism of action of Ephedra Herba Decoction against influenza A virus based on network pharmacology

Objective To predict the main bioactive components and potential mechanisms of Ephedra Herba Decoction(Chinese Pinyin abbreviated as MHT)in treating influenza A virus(IAV)based on network pharmacology.Methods Multiple online databases were used to search and screen out the active components from MHT,the related targets of active components of MHT and the genes related to IAV.Search the corresponding genes name of target through UniProt database.Cytoscape 3.7.2 was used to construct the drug-component-target network diagram.Venn diagram was used to screen the intersection genes of the active components corresponding to the target and disease-related genes,and the intersection genes were imported into the STRING Database Online platform to obtain the protein-protein interaction(PPI)network.Then,the PPI network was imported into Cytoscape 3.7.2 to obtain the core targets.Finally,Wei Sheng Xing(http://www.bioinformatics.com.cn/)was used to do GO function enrichment analysis and KEGG signaling pathway enrichment analysis for the intersection genes,and the results of GO and KEGG were visualized.Results A total of 116 active components and 253 potential targets were screened from MHT.Quercetin,Kaempferol,and Luteolin are the main active components,and AKT1,TNF,TP53,IL6,and JUN are the core targets of 253 potential targets.There are 2906 targets of influenza A,including 121 intersection genes.Enrichment analysis showed that there were 131 entries of molecular function,89 entries of cell component,1645 entries of biological process and 195 entries of signaling pathway.PI3K-Akt,MAPK,and JAK-STAT were the main signaling pathways.Conclusion MHT plays an important role in the prevention and treatment of influenza A by acting on multiple targets and multiple signaling pathways.